Objective To investigate whether hemospermia is involved in the production of antisperm antibodies (ASA). Methods A total of 40 males with hemospermia were enrolled in this study. These males were divided into 3 groups according to their fertile status: 18 cases were patients who visited the infertility clinic (group A), 8 had fertility (group B), and 14 did not know their fertile status (group C). Semen samples were collected and ASA levels were detected by the direct immunobead test (D-IBT). Leukocytes in semen were assessed by the peroxidase assay. Results None of these males with hemospermia were found to display significant levels of ASA in the 3 groups. The number of cases who had increased levels of leukocytes in the semen samples of groups A, B and C were 15 (83.3%), 6 (75.0%) and 12 (85.7%), respectively. Conclusion Hemospermia is not associated with the production of ASA. It is worth noting that the leukocyte levels are increased in semen of males with hemospermia.